Avaliação do transplante de medula óssea alogênico por meio do estudo de regiões de repetições seqüenciais no genoma humano (VNTRs e STRs)

Os objetivos deste estudo foram estabelecer um protocolo para a análise de minissatélites ou VNTRs e microssatélites ou STRs em pacientes que se submeteram ao TMO alogênico; verificar a validade da metodologia e dos loci estudados e avaliar o tipo de recuperação do paciente. Foram analisados o DNA do paciente anterior e posterior ao transplante de 14 indivíduos e dos respectivos doadores. Amplificações por PCR de seis loci: D1S80, SE33, HumTH01, 33.6, HumARA e HumTPO foram realizadas. Os produtos amplificados foram separados por eletro­forese vertical em gel de poliacrilamida, e os fragmentos visualizados por coloração pela prata. Esse procedimento mostrou ser válido na verificação da recuperação alogênica, autóloga e provavelmente na quimérica. da somatória dos loci estudados, 63,1% apresentaram resultados possíveis de serem avaliados e, desses, 19,0% mostraram resultado informativo, 13,1% parcialmente informativo e 31,0% não informativo. Os 36,9% restantes não foram possíveis de avaliação. Dos loci avaliados, o que demostrou maior índice de resultado informativo foi o SE33, parcialmente informativo o HumTPO e não informativo o HumTH01, sendo o locus 33.6 o que mais apresentou resultados não possíveis de serem avaliados. Por outro lado, determinou-se a recuperação do paciente posterior ao transplante em 71,4% dos indivíduos, sendo que, desses, 90% apresentaram recuperação alogênica e 10% recuperação autóloga.

O Modelo de Ising inomogêneo: uma interrupção contínua entre as redes quadrada e triangular.

The ferromagnetic and antiferromagnetic Ising model on a two dimensional inhomogeneous lattice characterized by two exchange constants (J1 and J2) is investigated. The lattice allows, in a continuous manner, the interpolation between the uniforme square (J2 = 0) and triangular (J2 = J1) lattices. By performing Monte Carlo simulation using the sequential Metropolis algorithm, we calculate the magnetization and the magnetic susceptibility on lattices of differents sizes. Applying the finite size scaling method through a data colappse, we obtained the critical temperatures as well as the critical exponents of the model for several values of the parameter α = J2 J1 in the [0, 1] range. The ferromagnetic case shows a linear increasing behavior of the critical temperature Tc for increasing values of α. Inwhich concerns the antiferromagnetic system, we observe a linear (decreasing) behavior of Tc, only for small values of α; in the range [0.6, 1], where frustrations effects are more pronunciated, the critical temperature Tc decays more quickly, possibly in a non-linear way, to the limiting value Tc = 0, cor-responding to the homogeneous fully frustrated antiferromagnetic triangular case.

Effect of lentil tannins on albumin hydrolysis by trypsin

Os taninos da casca da semente de lentilha foram extraídos e purificados, levados à interação com albumina isolada de lentilha e com caseína; e estudados por turbidimetria. As interações da albumina e caseína com taninos purificados, a várias relações tanino-proteína, mostraram ser independente e dependente do pH, respectivamente. Hidrólise in vitro com tripsina das proteínas sem taninos indicou que o aquecimento a 99°C/15 min reduzia a susceptibilidade da albumina e aumentava a da caseína à tripsina. A influência de diferentes relações tanino:proteína (1:40; 1:20; 1:5; 1:2,5) na hidrólise mostrou maior inibição para caseína que para albumina de lentilha, independente de aquecimento. Após aquecimento ambas proteínas foram mais hidrolizadas para qualquer das relações tanino proteínas estudadas. A eletroforese em gel de poliacrilamida-dodecilsulfato de sódio do transcurso da hidrólise da interação tanino-albumina nativa mostra a dependência da relação tanino:proteína.

Caracterização e hidrólise in vitro da globulina principal de grão-de-bico (Cicer arietinum L.), var. IAC-Marrocos

No presente estudo procedeu-se ao isolamento e caracterização da fração globulina majoritária (11 S) de grão-de-bico, var. IAC-Marrocos. A globulina majoritária extraída foi isolada por cromatografia de filtração em gel e de troca-iônica mostrando apenas uma banda de proteína na eletroforese em gel de poliacrilamida. A globulina majoritária, após passagem em coluna de Sephadex, revelou duas bandas protéicas de 55 e 52,5kDa e três bandas menores em gel de poliacrilamida dodecilsulfato de sódio. Na presença de 2-mercaptoetanol 6 polipeptídios na faixa de 18 a 42kDa foram revelados na eletroforese. A globulina isolada foi submetida à ação da tripsina e quimotripsina onde a forma nativa mostrou-se resistente à ação enzimática enquanto o aquecimento (96 e 121°C/15min) não foi suficiente para aumentar a susceptibilidade à hidrólise, significativamente. Adição de NaCl 0,3M levou a um aumento da estabilidade estrutural com menor susceptibilidade à digestão proteolítica, fato em parte perdido com o aquecimento. As hidrólises foram acompanhadas por eletroforese em gel de poliacrilamida dodecilsulfato de sódio.

Effect of chickpea (Cicer arietinum L.) germination on the major globulin content and in vitro digestibility

A germinação das sementes de grão-de-bico foi acompanhada por um período de 6 dias, no qual pequenas variações nos teores de nitrogênio e globulina total foram registradas. A globulina majoritária (tipo 11 S) apresentou maiores variações após o quarto dia de germinação. A natureza e distribuição da fração globulina majoritária isolada na cromatografia em Sepharose CL-6B mostrou pequenas modificações ao final do período de germinação. A eletroforese em gel de poliacrilamida com dodecilssulfato de sódio do pico eluído na cromatografia em Sepharose CL-6B demonstra modificações nas bandas de proteínas entre os pesos moleculares de 20 e 30 kDa e acima de 60 kDa, indicando degradação protéica durante o período. Atividade proteolítica foi detectada na fração albumina da semente que aumentou até o quarto dia, seguido de queda até o sexto dia de germinação, quando da utilização de globulina total isolada da semente e caseína como substratos. Farinha de grão-de-bico, frações albumina e globulina total isoladas não apresentaram aumento na digestibilidade in vitro; entretanto, a fração globulina majoritária isolada foi mais suscetível à hidrólise após germinação.

Propriedades críticas do modelo z(4) em sistemas bi e tridimensionais

A real space renormalization group method is used to investigate the criticality (phase diagrams, critical expoentes and universality classes) of Z(4) model in two and three dimensions. The values of the interaction parameters are chosen in such a way as to cover the complete phase diagrams of the model, which presents the following phases: (i) Paramagnetic (P); (ii) Ferromagnetic (F); (iii) Antiferromagnetic (AF); (iv) Intermediate Ferromagnetic (IF) and Intermediate Antiferromagnetic (IAF). In the hierarquical lattices, generated by renormalization the phase diagrams are exact. It is also possible to obtain approximated results for square and simple cubic lattices. In the bidimensional case a self-dual lattice is used and the resulting phase diagram reproduces all the exact results known for the square lattice. The Migdal-Kadanoff transformation is applied to the three dimensional case and the additional phases previously suggested by Ditzian et al, are not found

Efeitos das inomogeneidades da matéria em Cosmologias Aceleradas

The recent observational advances of Astronomy and a more consistent theoretical framework turned Cosmology in one of the most exciting frontiers of contemporary science. In this thesis, homogeneous and inhomogeneous Universe models containing dark matter and different kinds of dark energy are confronted with recent observational data. Initially, we analyze constraints from the existence of old high redshift objects, Supernovas type Ia and the gas mass fraction of galaxy clusters for 2 distinct classes of homogeneous and isotropic models: decaying vacuum and X(z)CDM cosmologies. By considering the quasar APM 08279+5255 at z = 3.91 with age between 2-3 Gyr, we obtain 0,2 < OM < 0,4 while for the j3 parameter which quantifies the contribution of A( t) is restricted to the intervalO, 07 < j3 < 0,32 thereby implying that the minimal age of the Universe amounts to 13.4 Gyr. A lower limit to the quasar formation redshift (zJ > 5,11) was also obtained. Our analyzes including flat, closed and hyperbolic models show that there is no an age crisis for this kind of decaying A( t) scenario. Tests from SN e Ia and gas mass fraction data were realized for flat X(z)CDM models. For an equation of state, úJ(z) = úJo + úJIZ, the best fit is úJo = -1,25, úJl = 1,3 and OM = 0,26, whereas for models with úJ(z) = úJo+úJlz/(l+z), we obtainúJo = -1,4, úJl = 2,57 and OM = 0,26. In another line of development, we have discussed the influence of the observed inhomogeneities by considering the Zeldovich-Kantowski-DyerRoeder (ZKDR) angular diameter distance. By applying the statistical X2 method to a sample of angular diameter for compact radio sources, the best fit to the cosmological parameters for XCDM models are OM = O, 26,úJ = -1,03 and a = 0,9, where úJ and a are the equation of state and the smoothness parameters, respectively. Such results are compatible with a phantom energy component (úJ < -1). The possible bidimensional spaces associated to the plane (a , OM) were restricted by using data from SNe Ia and gas mass fraction of galaxy clusters. For Supernovas the parameters are restricted to the interval 0,32 < OM < 0,5(20") and 0,32 < a < 1,0(20"), while to the gas mass fraction we find 0,18 < OM < 0,32(20") with alI alIowed values of a. For a joint analysis involving Supernovas and gas mass fraction data we obtained 0,18 < OM < 0,38(20"). In general grounds, the present study suggests that the influence of the cosmological inhomogeneities in the matter distribution need to be considered with more detail in the analyses of the observational tests. Further, the analytical treatment based on the ZKDR distance may give non-negligible corrections to the so-calIed background tests of FRW type cosmologies

Floração precoce em cana-de-açúcar - um estudo utilizando ferramentas de análise in silico e proteômica

Sugarcane is one of the most important products of the world and Brazil is responsible for 25 % of the world production. One problem of this culture at northeast of Brazil is the early flowering. In our laboratory, it has been made before four subtractive libraries using early and late flowering genotypes in order to identify messages related to the flowering process. In this work, two cDNAs were chosen to make in silico analysis and overexpression constructs. Another approach to understand the flowering process in sugarcane was to use proteomic tools. First, the protocol for protein extraction using apical meristem was set up. After that, these proteins were separated on two bidimensional gels. It was possible to observe some difference for some regions of these gels as well as some proteins that can be found in all conditions. The next step, spots will be isolated and sequence on MS spectrometry in order to understand this physiological process in sugarcane

Análise molecular dos éxons 8 a 11 do gene da p53 em amostras de câncer de colo de útero no Rio Grande do Norte

Mutations on TP53 gene are common in human cancer but not in cervical cancer where they are rarely found and the inactivation and degradation of p53 protein are attributed to the action of E6 viral oncogene from high risk human papillomavirus (HPV). Analysis of cervical cancer cell lines suggests that HPV negative samples shows mutation on TP53, but clinical approaches didn t confirmed this hypothesis. However, in most TP53 mutations studies on cervical cancer, only the exons 5 to 8 were analyzed. Approximately 90% of mutations described are on this region. Recent studies on several cancer suggests that mutation frequency in the other exons must be considered. The aim of this work was to verify whether mutations on coding and non-coding regions occur in cancer tissue from cervical cancer in patients from Rio Grande do Norte using Denaturing Gradient Gel Electrophoresis (DGGE) as screening tool. Exons 8 to 11 were analyzed including some introns from 80 tumor samples and 8 peripheral blood samples from healthy women. DNA were submitted to PCR using primers with GC clamp on the end of one of them. The results were observed for each region after DGGE and silver staining. It was observed no amplified fragment with different migration profile from those obtained from DNA of peripheral blood. These results agree with those from literature where TP53 mutations in cervical cancer have been described in a very low frequency

Desenvolvimento de sistemas farmacêuticos emulsionados para veiculação gênica

Broadly speaking, the concept of gene therapy involves the transfer of a genetic material into a cell, tissue, or organ in order to cure a disease or at least improve the clinical status of a patient. Making it simple, gene therapy consists in the insertion of functional genes into cells containing defective genes by substituting, complementing or inhibiting them. The achievement of a foreigner DNA expression into a population of cells requires its transfer to the target. Therefore, it is a key issue to create systems able to transfer and protect the DNA until it reaches the target, the vectors. The disadvantages related to the use of viral vectors have encouraged efforts to develop emulsions as non-viral vectors. In fact, they are easily produced, present controllable stability and enable transfection. The aim of this work was to develop an emulsion for gene therapy and evaluate its ability to compact nucleic acids by the development of a complex with the plasmid pIRES2-EGFP. The first step was to determine the Hydrophilic Lipophilic Balance (HLB) of the Captex® 355 (oily internal phase of the emulsion) through long and short term stability assays. Based on the results, emulsions composed of Captex® 355, Tween 20® and Span 60® with 10.7 HLB were produced by three different methods: phase inversion, spontaneous emulsification and sonication. The results showed that the lowest diameter and best stability of the emulsions were achieved by the sonication method. The cationic emulsions were made by adding DOTAP to the basic emulsion. Its association with pIRES2-EGFP was evaluated by electrophoresis. Several rates of emulsion and DNA were evaluated and the results showed that 100% of the complex was formed when the rate DOTAP/DNA(nmol/µg) was 130. In conclusion, the overall results show the ability of the proposed emulsion to compact pIRES2-EGFP, which is a requirement to a successful transfection. Therefore, such formulation may be considered a promising candidate for gene therapy

Estudo integrado do potencial fitorremediador da Eichhornia crassipes em ambientes naturais e sua utilização para obtenção de extratos proteicos

This study aimed to analyze the phytoremediation potential of Eichhornia crassipes in natural environments, optimize the extraction process of crude protein from plant tissue and, obtain and characterize this process in order to determine its viability of use instead of the protein sources of animal and/or human feed. For this, it has been determined in Apodi/Mossoró river water the concentration of ammonium ions, nitrite, nitrate, calcium, magnesium, potassium, iron, copper, manganese, zinc, nickel, cobalt, sodium, aluminum, cádmium, lead, and total chromium; It was determined in plant tissue of aquatic macrophytes of Eichhornia crassipes species present in Apodi/Mossoró River the moisture content, ash, calcium, magnesium, potassium, iron, copper, manganese, zinc, nickel, cobalt, sodium, aluminum, cadmium, lead, total chromium, total nitrogen and crude protein. It was also determined the translocation factor and bioaccumulation of all the quantified elements; It was developed and optimized the extraction procedure of crude protein based on the isoelectric method and a factorial design 24 with repetition; It was extracted and characterized the extract obtained by determining the moisture content, ash, magnesium, potassium, iron, copper, manganese, zinc, nickel, cobalt, sodium, cadmium, total nitrogen and crude protein. And finally, it was also characterized the protein extract using Thermogravimetric Analysis (TG), Derived Thermogravimetric (DTG), Differential Scanning Calorimetry (DSC), Infrared Spectroscopy (FT-IR) and jelly-like electrophoresis of polyacrylamide (SDS -PAGE) to assess the their molecular weights/mass. Thus, from the results obtained for the translocation and bioaccumulation factors was found that the same can be used as phytoremediation agent in natural environments of all quantified elements. It was also found that the developed method of extraction and protein precipitation was satisfactory for the purpose of the work, which gave the best conditions of extraction and precipitation of proteins as: pH extraction equal to 13.0, extraction temperature equals 60 ° C, reaction time equals to 30 minutes, and pH precipitation equals to 4.0. As for the extract obtained, the total nitrogen and crude protein were quantified higher than those found in the plant, increasing the crude protein content approximately 116.88% in relation to the quantified contente in the vegetal tissue of macrophyte. The levels of nickel and cadmium were the unique that were found below the detection limit of used the equipment. The electrophoretic analysis allowed us to observe that the protein extract obtained is composed of low polypeptide chains by the molecular and phytochelatins, with 6 and 15 kDa bands. Analysis of TG, DTG, DSC and FT-IR showed similarities in protein content of the obtained extracts based on different collection points and 9 parts of the plant under study, as well as commercial soy protein and casein. Finally, based on all these findings, it was concluded that the obtained extract in this work can be used instead of the protein sources of animal feed should, before that, test its digestibility. As human supplementation, it is necessary to conduct more tests associated with the optimization process in the sense of removing undesirable components and constant monitoring of the water body and the raw material used

Bolton freeware: aplicativo de avaliação de discrepância de massa dentária desenvolvido pelo programa de pós-graduação em odontologia/UFRN

The calculation of tooth mass discrepancy, essential for good planning and a proper orthodontic finishing, when performed manually, besides being laborious, requires considerable time consumption. The aim of this study was to develop and test Bolton Freeware, a software for analysis of the tooth mass discrepancy of Bolton, aiming to minimize the consumption of time in a less onerous way. The digital analysis of the software was done by means of two-dimensional scanning of plaster study models and compared to manual evaluation (gold standard), using 75 pairs of stone plaster study models divided into two groups according to the magnitude of the Curve of Spee (group I from 0 to 2 mm, group II greater than 2 to 3mm). All the models had permanent dentition and were in perfect condition. The manual evaluation was performed with a digital caliper and a calculator, and the time required to perform the analysis for both methods was recorded and compared. In addition, the software was evaluated by orthodontists regarding its use, by means of questionnaires developed specifically for this purpose. Calibration was performed prior to manual analysis, and excellent levels of inter-rater agreement were achieved, with ICC > 0.75 and r > 0.9 for total and anterior proportion. It was observed in the evaluation of error of the digital method that some teeth showed a significant systematic error, being the highest measured at 0.08 mm. The analysis of total tooth mass discrepancy performed by Bolton Freeware, for those cases in which the curve of Spee is mild and moderate, differ from manual analysis, on average, 0.09 mm and 0.07 mm respectively, for each tooth evaluated, with r> 0, 8 for total and anterior proportion. According to the specificity and sensitivity test, Bolton Freeware has an improved ability to detect true negatives, i.e. the presence of discrepancy. The Bolton analysis digitally performed was faster, with an average difference of time consumed to perform the analysis of Bolton between the two methods of approximately 6 minutes. Most experts interviewed (93%) approved the usability of the software

Perfil eletroforético e concentração de imunoglobulinas G (IgG) do soro sanguíneo de cabras Saanen com mastite experimental induzida por Staphylococcus aureus suplementadas com vitamina E

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Perfil eletroforético das proteínas séricas de serpentes Crotalus durissus terrificus (cascavel) criadas em cativeiro

As serpentes peçonhentas dos gêneros Bothrops e Crotalus têm sido mantidas em cativeiro visando à extração de venenos para a produção de imunobiológicos. O conhecimento da fisiologia desses animais e as alterações na concentração de proteínas e suas frações séricas são importantes para a identificação precoce de importantes enfermidades que cursam com estados de hipoproteinemia e hiperproteinemia. O objetivo do trabalho foi determinar a concentração de proteína total e o perfil eletroforético das proteínas séricas de serpentes Crotalus durissus terrificus (cascavel) criadas em cativeiro. Foram colhidas amostras de sangue da veia coccígea ventral de 21 serpentes adultas e sadias, divididas em dois grupos: Grupo 1 de 12 machos com peso médio de 588,89±193,55g, e Grupo 2 de nove fêmeas com peso médio de 708,33±194,04g. A proteína total sérica foi determinada pelo método de refratometria e a eletroforese em gel de agarose. Obtiveram-se valores da proteína total sérica (g/dL) de 4,51±0,50 para machos e de 4,82±0,72 para fêmeas, e para machos e fêmeas de 4,64±0,61. Foram identificadas pela eletroforese quatro frações protéicas (g/dL): albumina, a, b, g-globulinas e calculada a relação albumina:globulina. As serpentes fêmeas apresentaram maiores valores para as variáveis, albumina e para a relação albumina/globulina (AG) diferindo significativamente (P<0,05) do grupo de machos, porém sem significado clínico.

Evaluation of nutritional status in patients with endemic pemphigus foliaceus

A avaliação nutricional foi realizada em dezesseis doentes portadores de pênfigo foliáceo endêmico, sendo dez portadores da forma localizada da doença (Grupo G1) e seis portadores da forma disseminada da mesma (Grupo G2). Foram realizadas avaliações antropométricas (peso, altura, índice de Quetelét, prega cutânea tricipital, prega cutânea subescapular, circunferência braquial, circunferência muscular do braço, área do braço, área muscular do braço, área adiposa do braço) e laboratorial (eletroforese de proteínas séricas). Quanto aos parâmetros antropométricos, observaram-se as seguintes alterações: a circunferência do braço, área do braço, e área muscular do braço mostraram valores mais baixos nos doentes do grupo G2 que nos do grupo G1. O peso e a circunferência muscular do braço, por sua vez, mostraram tendência a valores mais baixos nos doentes do grupo G2 que nos do grupo G1. A eletroforese de proteínas revelou valores de albumina diminuídos em ambos os grupos, e menores nos doentes do grupo G2. Quanto às demais frações, com exceção das beta globulinas, não foram evidenciadas alterações. A análise global dos resultados permite concluir que os doentes com pênfigo foliáceo endêmico apresentam um quadro de desnutrição proteica, mas não calórica. Esta desnutrição mostrou-se mais acentuada no pênfigo foliáceo disseminado.